The differential roles of D-Pax2 variants in regulating Drosophila eye and bristle development

The ability to appropriately interact with the environment is crucial to an organism’s survival. The establishment of functional sensory systems, such as the bristles and eyes in Drosophila, is a critical event during the development of the organism. The transcription factor D Pax2 is involved in the differentiation of the shaft and glial cells in the developing bristle (Kavaler et al., Dev, 126:2261-2272, 1999) and of the cone and primary pigment cells in the developing eye (Fu and Noll, Genes Dev, 11:389-405, 1997). How D-Pax2 contributes to distinct differentiative pathways in different cell types is not known. Recent work by Anna Czechowski and Katherine Harmon (personal communication) identified a mutation in the D-Pax2 gene that introduced a stop codon at the end of exon 9, effectively truncating the protein. This mutation affects bristle, but not eye, development. We thus suspected regions after exon 9 are required for D-Pax2 function only in the bristles and may also be associated with alternative splicing of the D Pax2 transcript. We plan to assess the role of the carboxy terminal region of the protein by establishing transgenic lines bearing rescue constructs of D-Pax2 with either the complete coding sequence or with deletions of specific exons. To date, we have generated the first rescue construct bearing the complete coding region of the gene driven by a 3 KB upstream regulatory region of D-Pax2 and are currently generating transgenic fly lines with this construct.

The Pollination Biology and Mating System of a Peripheral Population of Witheringia solanacea (Solanaceae)

Pollinator visitation rates over the life of a flower are determined by pollinator abundance and floral longevity. If flowers are not visited frequently enough, pollen limitation may occur, favoring the evolution of self-compatibility (SC). In plant species with varying SC levels, central populations often are self-incompatible (SI) and peripheral populations are SC. Witheringia solanacea (Solanaceae) is a species that follows this trend with the exception of one population in the Monteverde Cloud Forest Reserve, which is peripheral yet SI. I investigated this population using multiple techniques including floral bagging, pollinator observations, microsatellite analysis, and floral longevity manipulations. My results confirmed the self-incompatibility of the Monteverde population and indicated low but perhaps adequate rates of pollinator visitation per flower per hour. I found reduced genetic diversity at Monteverde and gene flow occurring unidirectionally from San Luis (a central population) to Monteverde. In the greenhouse, there was more of an effect of male than female function on floral longevity, but the largest differences were environmental. Flowers stayed open substantially longer when cool, cloudy weather was simulated and shorter when conditions were hot and sunny. The results indicate that the Monteverde population of W. solanacea is SI because 1) it is unable to maximize its fitness due to gene flow from San Luis and its relatively recent colonization of the area and 2) pollen limitation may not be severe because of supplemental pollinator availability from other Witheringia species in the area and increased floral longevities due to cool and cloudy conditions.